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当前位置: 首页 > 文章 > 犬IFN-γ的原核表达及其对A型流感病毒H3N2亚型的抑制作用 动物医学进展 2016 (7) 1-6
Position: Home > Articles > Prokaryotic Expression of Canine IFN-γ and Analysis of Its Antiviral Activities to Canine Influenza Virus A H3N2 Progress in Veterinary Medicine 2016 (7) 1-6

犬IFN-γ的原核表达及其对A型流感病毒H3N2亚型的抑制作用

作  者：
秦海斌;朱海娟;朱骞;宋珍华;温海;贺星亮;高一龙
单  位：
公安部南京警犬研究所警犬技术公安部重点实验室
关键词：
犬;γ干扰素;原核表达;流感病毒
摘  要：
为研究犬的γ干扰素(IFN-γ)对犬流感病毒H3N2亚型的抑制作用,由经ConA诱导过的健康犬淋巴细胞中特异性地扩增犬IFN-γ(cIFN-γ)基因,构建重组原核表达载体pET-cIFN-γ,转化宿主菌BL21(DE3)中进行诱导表达,以MDCK-VSV法对纯化后产物进行抗病毒活性鉴定,并在MDCK细胞上测定对H3N2亚型犬流感病毒的抑制作用。结果表明,表达产物以包涵体形式存在,经变性、复性、纯化后,所制备犬IFN-γ的抗病毒活性为1.1×10~5 U/mg,重组犬IFN-γ稀释2~6倍后对犬流感病毒的增殖仍具有明显抑制作用。本研究成功表达了具备抗H3N2亚型流感病毒活性的犬IFN-γ,为犬流感病毒新型药物的开发奠定了基础。
译  名：
Prokaryotic Expression of Canine IFN-γ and Analysis of Its Antiviral Activities to Canine Influenza Virus A H3N2
作  者：
QIN Hai-bin;ZHU Hai-juan;ZHU Qian;SONG Zhen-hua;WEN Hai;HE Xing-liang;GAO Yi-long;Nanjing Policedog Research Institute of MPS,Policedog Technology Key Laboratory of MPS;
关键词：
canine;;interferon-γ;;prokaryotic expression;;Influenza virus
摘  要：
To prepare recombinant canine interferon gamma(IFN-γ)and study its antiviral effect to canine inflenza H3N2,the mature IFN-γgene was amplied from template NDA extracted from healthy canine's serum lymphocytes which were induced by ConA.IFN-γgene was insert into pET-32 ato construct pET-c IFN-γexpression vector.pET-c IFN-γwas tansformed into E.coli BL21,in which fused protein can be expressed by IPTG inducing.The expression product was purefied and the antiviral activity to H3N2 was tested with MDCK-VSV method.Results showed that the fusion protein size was 34 ku and existed in form of inclusion bodies.After refolding and purification,the recombinant protein still had high antiviral activity about 1.1×10~5 U/mg to VSV in MDCK.The dilution of 2~6 of IFN-γcould protect MDCK from the cytopathic effect caused by 100TCID_(50) of canine H3N2 virus.The results proved we successfully expressed the canine IFN-γin prokaryotic system and laid a good foundation for the development of new drugs anti-canine influenza virus.

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